Increase in anti-IFNα autoantibodies results in a decline in tissue infiltrations and inflammation. (A) Pathology scores of the salivary gland and pancreas of the Aire-deficient (KO, red) and control (HE, blue) rats across various ages (above) and correlation between anti-IFNα11 and pathology scores of all Aire-deficient animals with detectable autoantibodies (below). Pathology scores were calculated as described in Materials and methods (n = 4–8 for each age group and 25 and 22 KO animals for all pooled antibody-positive salivary gland and pancreas samples, respectively; data are combined from three or more individual experiments). P values are based on t tests (salivary gland) and the Fisher–Pitman permutation test (pancreas). P values are adjusted for multiple comparisons using the Holm–Bonferroni method. (B) Representative samples of H&E (HE) stained pancreatic and salivary gland sections of Aire-deficient (KO) and control (HE) rats (n = 4–7 for both groups of KO and HE rats, data are combined from three or more individual experiments). Black arrows highlight cellular infiltrations (100× magnification, scale bar 250 μm). Note that some of the H&E stainings are duplicated in Fig. S5. (C) Relative change in expression of ISGs after incubating cultured splenic cells with blood plasma from Aire-deficient rats as quantified using qPCR (n = 4 for both KO and HE rats, combined from two independent experiments). P values are based on t tests. P values are adjusted for multiple comparisons using the Holm–Bonferroni method. (D) The effect of Imiquimod versus vehicle (vaseline) on ear thickening in Aire-deficient (KO) and control (WT) rats determined as described in the Materials and methods (shown as mean value with SEM, n = 4 for both KO and WT rats, data obtained from a single experiment). P values are based on two-way ANOVA with post-hoc Wilcoxon signed-rank test for pairwise comparisons; *P < 0.05. (E) UMAP embedding of scRNA-seq data obtained from FACS-sorted CD45+ cells purified from the salivary glands of 7-mo-old Aire-deficient (KO) and control (HE) rats and colored by cell type. (F) Volcano plots showing changes in the expression of ISGs (red) in CD8+ T cells, macrophages, NK cells, and Treg cells from the salivary glands of 7-mo-old Aire-deficient and control rats. Data in E and F are obtained from a single experiment with four Aire-deficient (KO) and three control (HE) rats per group. In A and C, symbols indicate individual animals and horizontal lines with whiskers indicate mean with SEM. Fisher’s exact test was performed to quantify enrichment of ISGs among downregulated (LFC < −1, FDR < 0.05) genes. Statistical significance is indicated as follows: ****P < 1e-4.
Figure 7.

Increase in anti-IFNα autoantibodies results in a decline in tissue infiltrations and inflammation. (A) Pathology scores of the salivary gland and pancreas of the Aire-deficient (KO, red) and control (HE, blue) rats across various ages (above) and correlation between anti-IFNα11 and pathology scores of all Aire-deficient animals with detectable autoantibodies (below). Pathology scores were calculated as described in Materials and methods (n = 4–8 for each age group and 25 and 22 KO animals for all pooled antibody-positive salivary gland and pancreas samples, respectively; data are combined from three or more individual experiments). P values are based on t tests (salivary gland) and the Fisher–Pitman permutation test (pancreas). P values are adjusted for multiple comparisons using the Holm–Bonferroni method. (B) Representative samples of H&E (HE) stained pancreatic and salivary gland sections of Aire-deficient (KO) and control (HE) rats (n = 4–7 for both groups of KO and HE rats, data are combined from three or more individual experiments). Black arrows highlight cellular infiltrations (100× magnification, scale bar 250 μm). Note that some of the H&E stainings are duplicated in Fig. S5. (C) Relative change in expression of ISGs after incubating cultured splenic cells with blood plasma from Aire-deficient rats as quantified using qPCR (n = 4 for both KO and HE rats, combined from two independent experiments). P values are based on t tests. P values are adjusted for multiple comparisons using the Holm–Bonferroni method. (D) The effect of Imiquimod versus vehicle (vaseline) on ear thickening in Aire-deficient (KO) and control (WT) rats determined as described in the Materials and methods (shown as mean value with SEM, n = 4 for both KO and WT rats, data obtained from a single experiment). P values are based on two-way ANOVA with post-hoc Wilcoxon signed-rank test for pairwise comparisons; *P < 0.05. (E) UMAP embedding of scRNA-seq data obtained from FACS-sorted CD45+ cells purified from the salivary glands of 7-mo-old Aire-deficient (KO) and control (HE) rats and colored by cell type. (F) Volcano plots showing changes in the expression of ISGs (red) in CD8+ T cells, macrophages, NK cells, and Treg cells from the salivary glands of 7-mo-old Aire-deficient and control rats. Data in E and F are obtained from a single experiment with four Aire-deficient (KO) and three control (HE) rats per group. In A and C, symbols indicate individual animals and horizontal lines with whiskers indicate mean with SEM. Fisher’s exact test was performed to quantify enrichment of ISGs among downregulated (LFC < −1, FDR < 0.05) genes. Statistical significance is indicated as follows: ****P < 1e-4.

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